IP-10 antibodies and their uses

Abstract

The present invention provides isolated monoclonal antibodies, particularly human antibodies, that bind to IP-10 with high affinity, inhibit the binding of IP-10 to its receptor, inhibit IP-10-induced calcium flux and inhibit IP-10-induced cell migration. Nucleic acid molecules encoding the antibodies of the invention, expression vectors, host cells and methods for expressing the antibodies of the invention are also provided. Immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the antibodies of the invention are also provided. The invention also provides methods for inhibiting IP-10 activity using the antibodies of the invention, including methods for treating various inflammatory and autoimmune diseases.

1 . An isolated human monoclonal antibody, or an antigen-binding portion thereof, wherein the antibody specifically binds to human IP-10 and exhibits at least one of the following properties: 
 (a) inhibits binding of IP-10 to CXCR3;    (b) inhibits IP-10 induced calcium flux;    (c) inhibits IP-10 induced cell migration;    (d) cross-reacts with rhesus monkey IP-10;    (e) does not cross-react with mouse IP-10;    (f) does not cross-react with human MIG;    (g) does not cross-react with human ITAC.    
     
     
         2 . The antibody of  claim 1 , which is full-length antibody of an IgG1 or IgG4 isotype.  
     
     
         3 . The antibody of  claim 1 , which is an antibody fragment or a single chain antibody.  
     
     
         4 . An isolated monoclonal antibody, or an antigen-binding portion thereof, wherein the antibody specifically binds to IP-10 and comprises a heavy chain variable region that is the product of or derived from a human V H  germline gene selected from the group consisting of a human V H  3-33 gene, a human V H  3-30.3 gene, a human V H  5-51 gene and a human V H  4-61 gene.  
     
     
         5 . The antibody of  claim 4 , wherein the IP-10 is human IP-10.  
     
     
         6 . An isolated monoclonal antibody, or an antigen-binding portion thereof, wherein the antibody specifically binds to IP-10 and comprises a light chain variable region that is the product of or derived from a human V k  germline gene selected from the group consisting of a human V k  A27 gene, a human V k  L15 gene, a human V k  L6 gene and a human V k  L18 gene.  
     
     
         7 . The antibody of  claim 6 , wherein the IP-10 is human IP-10.  
     
     
         8 . An isolated monoclonal antibody, or an antigen-binding portion thereof, wherein the antibody specifically binds to IP-10 and comprises: 
 (a) a heavy chain variable region that is the product of or derived from a human V H  germline gene selected from the group consisting of a human V H  3-33 gene, a human V H  3-30.3 gene, a human V H  5-51 gene and a human V H  4-61 gene; and    (b) a light chain variable region that is the product of or derived from a human V k  germline gene selected from the group consisting of a human V k  A27 gene, a human V k  L15 gene, a human V k  L6 gene and a human V k  L18 gene.    
     
     
         9 . The antibody of  claim 8 , wherein the IP-10 is human IP-10.  
     
     
         10 . The antibody of  claim 8 , which comprises: 
 (a) a heavy chain variable region that is the product of or derived from a human V H  3-33 gene; and    (b) a light chain variable region that is the product of or derived from a human Vk germline gene selected from the group consisting of a human Vk A27 gene, a human Vk L15 gene and a human Vk L6 gene.    
     
     
         11 . The antibody of  claim 10 , which comprises a light chain variable region that is the product of or derived from a human Vk A27 gene.  
     
     
         12 . The antibody of  claim 10 , which comprises a light chain variable region that is the product of or derived from a human Vk L15 gene.  
     
     
         13 . The antibody of  claim 10 , which comprises a light chain variable region that is the product of or derived from a human Vk L6 gene.  
     
     
         14 . The antibody of  claim 8 , which comprises: 
 (a) a heavy chain variable region that is the product of or derived from a human V H  3-30.3 gene; and    (b) a light chain variable region that is the product of or derived from a human Vk L6 gene.    
     
     
         15 . The antibody of  claim 8 , which comprises: 
 (a) a heavy chain variable region that is the product of or derived from a human V H  5-51 gene; and    (b) a light chain variable region that is the product of or derived from a human Vk L18 gene.    
     
     
         16 . The antibody of  claim 8 , which comprises: 
 (a) a heavy chain variable region that is the product of or derived from a human V H  4-61 gene; and    (b) a light chain variable region that is the product of or derived from a human Vk A27 gene.    
     
     
         17 . An isolated monoclonal antibody, or antigen binding portion thereof, comprising: 
 a heavy chain variable region that comprises CDR1, CDR2, and CDR3 sequences; and    a light chain variable region that comprises CDR1, CDR2, and CDR3 sequences, wherein:    (a) the heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 24-34, and conservative modifications thereof,    (b) the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequence of SEQ ID NOs: 73-83, and conservative modifications thereof,    (c) the antibody specifically binds to IP-10, and    (d) the antibody exhibits at least one of the following functional properties: 
 (i) the antibody inhibits binding of IP-10 to CXCR3;  
 (ii) the antibody inhibits IP-10 induced calcium flux;  
 (iii) the antibody inhibits IP-10 induced cell migration;  
 (iv) the antibody cross-reacts with rhesus monkey IP-10;  
 (v) the antibody does not cross-react with mouse IP-10;  
 (vi) the antibody does not cross-react with human MIG;  
 (vii) the antibody does not cross-react with human ITAC.  
   
     
     
         18 . The antibody of  claim 17 , wherein the heavy chain variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 13-23, and conservative modifications thereof; and the light chain variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 62-72, and conservative modifications thereof.  
     
     
         19 . The antibody of  claim 18 , wherein the heavy chain variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 1-12, and conservative modifications thereof; and the light chain variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 51-61, and conservative modifications thereof.  
     
     
         20 . The antibody of  claim 17 , which is a human antibody.  
     
     
         21 . The antibody of  claim 17 , which is a humanized or chimeric antibody.  
     
     
         22 . An isolated monoclonal antibody, or antigen binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein: 
 (a) the heavy chain variable region comprises an amino acid sequence that is at least 80% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 35-46,    (b) the light chain variable region comprises an amino acid sequence that is at least 80% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs:84-94, 
 (c) the antibody specifically binds to IP-10, and  
 (d) the antibody exhibits at least one of the following functional properties: 
 (i) the antibody inhibits binding of IP-10 to CXCR3;  
 (ii) the antibody inhibits IP-10 induced calcium flux;  
 (iii) the antibody inhibits IP-10 induced cell migration;  
 (iv) the antibody cross-reacts with rhesus monkey IP-10;  
 (v) the antibody does not cross-react with mouse IP-10;  
 (vi) the antibody does not cross-react with human MIG;  
 (vii) the antibody does not cross-react with human ITAC.  
 
   
     
     
         23 . The antibody of  claim 22 , which is a human antibody.  
     
     
         24 . The antibody of  claim 22 , which is a humanized or chimeric antibody.  
     
     
         25 . An isolated monoclonal antibody, or antigen binding portion thereof, comprising: 
 (a) a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-12;    (b) a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-23;    (c) a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-34;    (d) a light chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 51-61;    (e) a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 62-72; and    (f) a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 73-83;    wherein the antibody specifically binds IP-10.    
     
     
         26 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 1;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 13;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 24;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 51;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 62; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 73.    
     
     
         27 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 2;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 14;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 25;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 52;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 63; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 74.    
     
     
         28 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 3;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 15;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 26;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 53;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 64; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 75.    
     
     
         29 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 4;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 16;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 27;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 54;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 65; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 76.    
     
     
         30 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 5;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 17;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 28;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 55;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 66; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 77.    
     
     
         31 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 6;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 18;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 29;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 56;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 67; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 78.    
     
     
         32 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 7;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 19;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 30;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 57;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 68; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 79.    
     
     
         33 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 8;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 20;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 31;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 58;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 69; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 80.    
     
     
         34 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 9;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 21;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 32;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 59;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 70; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 81.    
     
     
         35 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 10 or 11;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 22;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 33;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 60;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 71; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 82.    
     
     
         36 . The antibody of  claim 25 , which comprises: 
 (a) a heavy chain variable region CDR1 comprising SEQ ID NO: 12;    (b) a heavy chain variable region CDR2 comprising SEQ ID NO: 23;    (c) a heavy chain variable region CDR3 comprising SEQ ID NO: 34;    (d) a light chain variable region CDR1 comprising SEQ ID NO: 61;    (e) a light chain variable region CDR2 comprising SEQ ID NO: 72; and    (f) a light chain variable region CDR3 comprising SEQ ID NO: 83.    
     
     
         37 . An isolated monoclonal antibody, or antigen binding portion thereof comprising: 
 (a) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 35-46; and    (b) a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 84-94;    wherein the antibody specifically binds IP-10.    
     
     
         38 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 35; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 84.    
     
     
         39 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 36; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 85.    
     
     
         40 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 37; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 86.    
     
     
         41 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 38; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 87.    
     
     
         42 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 39; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 88.    
     
     
         43 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 40; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 89.    
     
     
         44 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 41; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 90.    
     
     
         45 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 42; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 91.    
     
     
         46 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 43; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 92.    
     
     
         47 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 44 or 45; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 93.    
     
     
         48 . The antibody of  claim 37 , which comprises: 
 (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 46; and    (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO: 94.    
     
     
         49 . An isolated human antibody, or antigen-binding portion thereof, that competes for binding to IP-10 with the antibody of  claim 1 .  
     
     
         50 . An isolated human antibody, or antigen-binding portion thereof, that competes for binding to IP-10 with the antibody of  claim 25 .  
     
     
         51 . An isolated human antibody, or antigen-binding portion thereof, that competes for binding to IP-10 with the antibody of  claim 37 .  
     
     
         52 . A composition comprising the antibody, or antigen-binding portion thereof, of  claim 1 , and a pharmaceutically acceptable carrier.  
     
     
         53 . An immunoconjugate comprising the antibody, or antigen-binding portion thereof, of  claim 1 , linked to a therapeutic agent.  
     
     
         54 . A composition comprising the immunoconjugate of  claim 53  and a pharmaceutically acceptable carrier.  
     
     
         55 . A bispecific molecule comprising the antibody, or antigen-binding portion thereof, of  claim 1 , linked to a second functional moiety having a different binding specificity than said antibody, or antigen binding portion thereof.  
     
     
         56 . A composition comprising the bispecific molecule of  claim 55 , and a pharmaceutically acceptable carrier.  
     
     
         57 . An isolated nucleic acid molecule encoding the antibody, or antigen-binding portion thereof, of  claim 1 .  
     
     
         58 . An expression vector comprising the nucleic acid molecule of  claim 57 .  
     
     
         59 . A host cell comprising the expression vector of  claim 58 .  
     
     
         60 . A transgenic mouse comprising human immunoglobulin heavy and light chain transgenes, wherein the mouse expresses the antibody of  claim 1 .  
     
     
         61 . A hybridoma prepared from the mouse of  claim 60 , wherein the hybridoma produces said antibody.  
     
     
         62 . A method of inhibiting an inflammatory or autoimmune response mediated by activated T cells or NK cells comprising contacting the T cells or NK cells with the antibody, or antigen-binding portion thereof, of  claim 1 , such that the inflammatory or autoimmune response is inhibited.  
     
     
         63 . A method of treating an inflammatory or autoimmune disease in a subject in need of treatment comprising administering to the subject the antibody, or antigen-binding portion thereof, of  claim 1  such that the inflammatory or autoimmune disease in the subject is treated.  
     
     
         64 . A method of treating a viral or bacterial infection involving unwanted IP-10 activity in a subject in need of treatment comprising administering to the subject the antibody, or antigen-binding portion thereof, of  claim 1 , such that the viral or bacterial infection in the subject is treated.  
     
     
         65 . A method for preparing an anti-IP-10 antibody comprising: 
 (a) providing: (i) a heavy chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs: 1-12, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs: 13-23; and a CDR3 sequence that is selected from the group consisting of SEQ ID NOs: 24-34; or (ii) a light chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs: 51-61, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs: 62-72 and a CDR3 sequence that is selected from the gropu consisting of SEQ ID NOs: 73-83;    (b) altering at least one amino acid residue within at least one variable region antibody sequence, said sequence being selected from the heavy chain variable region antibody sequence and the light chain variable region antibody sequence, to create at least one altered antibody sequence; and    (c) expressing the altered antibody sequence as a protein.